Naturally Derived Phenethyl Isothiocyanate Modulates Induction of Oxidative Stress via Its N -Acetylated Cysteine Conjugated form in Malignant Melanoma.
Sotiris KyriakouNikoletta DemosthenousTom AmeryKyle J StewartPaul G WinyardRodrigo FrancoAglaia PappaMihalis I PanayiotidisPublished in: Antioxidants (Basel, Switzerland) (2024)
Phenethyl isothiocyanate (PEITC) is a secondary metabolic product yielded upon the hydrolysis of gluconasturtiin and it is highly accumulated in the flowers of watercress. The aim of the current study was to assess the role of a naturally derived PEITC-enriched extract in the induction of oxidative stress and to evaluate its anti-melanoma potency through the regulation of its metabolism with the concurrent production of the N -acetyl cysteine conjugated by-product. For this purpose, an in vitro melanoma model was utilized consisting of human primary (A375) cells as well as metastatic (COLO-679) malignant melanoma cells together with non-tumorigenic immortalized keratinocytes (HaCaT). Cytotoxicity was assessed via the Alamar Blue assay whereas the antioxidant/prooxidant activity of PEITC was determined via spectrophotometric assays. Finally, kinetic characterization of the end-product of PEITC metabolism was monitored via UPLC coupled to a tandem mass spectrometry (MS/MS). Our results indicate that although PhEF showed very minor antioxidant activity in a cell-free system, in a cell-based system, it can modulate the activity of key enzyme(s) involved in cellular antioxidant defense mechanism(s). In addition, we have shown that PhEF induces lipid and protein oxidation in a concentration-dependent manner, while its cytotoxicity is not only dependent on PEITC itself but also on its N -acetylated cysteine conjugated form.
Keyphrases
- oxidative stress
- induced apoptosis
- tandem mass spectrometry
- simultaneous determination
- cell free
- ultra high performance liquid chromatography
- high performance liquid chromatography
- photodynamic therapy
- ms ms
- diabetic rats
- liquid chromatography tandem mass spectrometry
- liquid chromatography
- fluorescent probe
- ischemia reperfusion injury
- high throughput
- dna damage
- living cells
- endothelial cells
- anti inflammatory
- gas chromatography
- squamous cell carcinoma
- high resolution
- single cell
- cell cycle arrest
- mass spectrometry
- cell therapy
- small molecule
- skin cancer
- fatty acid
- hydrogen peroxide
- stem cells
- induced pluripotent stem cells
- mesenchymal stem cells
- cell proliferation
- endoplasmic reticulum stress
- protein protein
- circulating tumor cells